Isolation, Purification and Characterization of Recombinantα-Galactosidase from Rice

doi:10.7506/spkx1002-6630-201021068

FOOD SCIENCE ›› 2010, Vol. 31 ›› Issue (21): 304-307.doi: 10.7506/spkx1002-6630-201021068

• Bioengineering • Previous Articles Next Articles

Isolation, Purification and Characterization of Recombinantα-Galactosidase from Rice

LI Su-hong1，ZHU Min-peng1，LI Tuo-ping2,*

1. College of Engineering and Technology, Shenyang Normal University, Shenyang 110034, China；
2. College of Light Industry, Liaoning University, Shenyang 110036, China

Received:2010-06-29 Online:2010-11-15 Published:2010-12-29
Contact: LI Tuo-ping2 E-mail:ltp0401@126.com

Abstract

Abstract:

A crude enzyme solution rich in recombinant riceα-galactosidase from the engineering strain E. coli pET-32a (+)- Gal/ Origami DE obtained from our laboratory was prepared and purified on a Ni-Sepharose affinity column. The purifiedα- galactosidase showed a single protein band in SDS-PAGE with a molecular mass of 59 kD. This enzyme exhibited the highest activity at the conditions of pH 5.0 and 45 ℃. Meanwhile, it showed the best stability at pH 4.0－7.0 and 0－20 ℃. The kinetic parameters Km and Vmax were 0.78 mmol/L and 10.16 mmol/(mg·min), respectively. Most of metal and organic ions did not affect the activity of this enzyme. However, Ag+, Hg2+ and pCMB (p-chloromercuribenzoic acid) significantly decreased its activity by 84%, 87% and 92%, respectively.

Key words: recombinant, α-galactosidase, purification, enzymatic characterization

CLC Number:

TS201.2

LI Su-hong1，ZHU Min-peng1，LI Tuo-ping2,*. Isolation, Purification and Characterization of Recombinantα-Galactosidase from Rice[J]. FOOD SCIENCE, 2010, 31(21): 304-307.

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Isolation, Purification and Characterization of Recombinantα-Galactosidase from Rice

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